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目的观察脂多糖(lipopolysac charide,LPS)作用下大鼠肺泡Ⅱ型上皮(typeⅡalveol arepith elial,AT-Ⅱ)细胞过氧化物酶增殖体激活受体γ(PPARγ)的表达变化情况。方法通过酶消化分离和免疫黏附法纯化原代培养AT-Ⅱ细胞,并进行碱性磷酸酶、肺表面活性物质相关蛋白A(SP-A)和电镜鉴定。采用RT-PCR法检测细胞经LPS作用后PPARγ、TNF-α、SP-AmRNA的改变,Westernblot法检测LPS作用后细胞PPARγ表达,ELISA法检测TNF-α蛋白表达。结果经碱性磷酸酶、SP-A和电镜鉴定,原代培养AT-Ⅱ细胞成功;在致炎因子LPS作用下,PPARγmRNA和蛋白表达均下降,这一变化伴随炎性因子TNF-α的升高。结论在LPS作用下,AT-Ⅱ细胞内PPARγ表达降低,提示PPARγ参与炎症反应。
Objective To observe the changes of peroxisome proliferator - activated receptor γ (PPARγ) expression in type Ⅱ alveol arepith elial (AT - Ⅱ) cells induced by lipopolysaccharide (LPS). Methods Primary cultured AT-Ⅱ cells were purified by enzymatic digestion and immunoadsorption. Alkaline phosphatase, pulmonary surfactant-associated protein A (SP-A) and electron microscopy were used to identify AT-Ⅱ cells. The changes of PPARγ, TNF-α and SP-AmRNA were detected by RT-PCR, the expression of PPARγ was detected by Western blot, and the expression of TNF-α was detected by ELISA. Results The primary culture of AT-Ⅱ cells was identified by alkaline phosphatase, SP-A and electron microscopy. The expression of PPARγmRNA and protein were decreased under the action of pro-inflammatory factor LPS. This change was associated with the increase of TNF-α high. Conclusion Under the action of LPS, the expression of PPARγ in AT-Ⅱ cells is decreased, suggesting that PPARγ is involved in the inflammatory reaction.